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EGFR and HER2 activate rigidity sensing only on rigid matrices

Nature Materials volume 16, pages 775–781 (2017)Cite this article

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Abstract

Epidermal growth factor receptor (EGFR) interacts with integrins during cell spreading and motility, but little is known about the role of EGFR in these mechanosensing processes. Here we show, using two different cell lines, that in serum- and EGF-free conditions, EGFR or HER2 activity increase spreading and rigidity-sensing contractions on rigid, but not soft, substrates. Contractions peak after 15–20 min, but diminish by tenfold after 4 h. Addition of EGF at that point increases spreading and contractions, but this can be blocked by myosin-II inhibition. We further show that EGFR and HER2 are activated through phosphorylation by Src family kinases (SFK). On soft surfaces, neither EGFR inhibition nor EGF stimulation have any effect on cell motility. Thus, EGFR or HER2 can catalyse rigidity sensing after associating with nascent adhesions under rigidity-dependent tension downstream of SFK activity. This has broad implications for the roles of EGFR and HER2 in the absence of EGF both for normal and cancerous growth.

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Figure 1: Rigidity-sensing activity measured by local CUs number.
Figure 2: Ligand-free EGFR and HER2 activity affects local CUs.
Figure 3: pEGFR localizes to early adhesions on rigid substrates.
Figure 4: EGFR and HER2 affect local contractility through Src.
Figure 5: EGF activates local contraction activity.

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Acknowledgements

We thank all the members of the Sheetz lab for their help. Especially M. X. Yao for preparing EGFR and HER2 mutants supported by MBI. This work was funded by National Institutes of Health (NIH) grant ‘Analysis of 120 nm local contractions linked to rigidity sensing’ (1 R01 GM100282-01), National Institutes of Health (NIH) grant ‘Tropomyosin and tyrosine kinases in mechanics of cancer’ (5 R01 GM113022-02) and by the NIH Common Fund Nanomedicine programme (PN2 EY016586). H.W. was supported by a Marie Curie International Outgoing Fellowship within the Seventh European Commission Framework Programme (PIOF-GA-2012-332045). B.Y. was supported by the MBI in Singapore. M.P.S. was partially supported by the Mechanobiology Institute, National University of Singapore.

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Author notes

  1. Mayur Saxena and Shuaimin Liu: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biomedical Engineering, Columbia University, New York, New York 10027, USA

    Mayur Saxena

  2. Department of Mechanical Engineering, Columbia University, New York, New York 10027, USA

    Shuaimin Liu, Cynthia Hajal, Junqiang Hu & James Hone

  3. Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore

    Bo Yang, Rishita Changede & Michael P. Sheetz

  4. Department of Biological Sciences, Columbia University, New York, New York 10027, USA

    Haguy Wolfenson & Michael P. Sheetz

  5. Department of Genetics and Developmental Biology, The Ruth and Bruce Rappaport Faculty of Medicine, The Technion—Israel Institute of Technology, Haifa 31096, Israel

    Haguy Wolfenson

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Contributions

M.S., S.L., B.Y., R.C. and J.H., performed the experiments; S.L. wrote MATLAB codes for data analysis; M.S., S.L., B.Y., C.H. and R.C. analysed the data; H.W., J.H., and M.P.S. designed the study; M.S., S.L., H.W., J.H. and M.P.S. wrote and prepared the manuscript.

Corresponding authors

Correspondence to Haguy Wolfenson, James Hone or Michael P. Sheetz.

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The authors declare no competing financial interests.

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Saxena, M., Liu, S., Yang, B. et al. EGFR and HER2 activate rigidity sensing only on rigid matrices. Nature Mater 16, 775–781 (2017). https://doi.org/10.1038/nmat4893

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